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大西洋鳕鱼(Gadus morhua)中五个干扰素调节因子转录本(Irf4a、Irf4b、Irf7、Irf8、Irf10)的特征及表达分析

Characterization and expression analyses of five interferon regulatory factor transcripts (Irf4a, Irf4b, Irf7, Irf8, Irf10) in Atlantic cod (Gadus morhua).

作者信息

Inkpen Sabrina M, Hori Tiago S, Gamperl A Kurt, Nash Gordon W, Rise Matthew L

机构信息

Department of Ocean Sciences, Memorial University of Newfoundland, NL, A1C 5S7, Canada.

出版信息

Fish Shellfish Immunol. 2015 May;44(1):365-81. doi: 10.1016/j.fsi.2015.02.032. Epub 2015 Feb 28.

Abstract

The interferon regulatory factor (IRF) family of genes encodes a group of transcription factors that have important roles not only in regulating the expression of Type I interferons (IFNs) and other genes in the IFN pathway, but also in growth, development and the regulation of oncogenesis. In this study, several IRF family members (Irf4a, Irf4b, Irf7, Irf8, Irf10) in Atlantic cod (Gadus morhua) were characterized at the cDNA and putative amino acid levels, allowing for phylogenetic analysis of these proteins in teleost fish, as well as the development of gene-specific primers used in RT-PCR and quantitative PCR (QPCR) analyses. Two Atlantic cod Irf10 splice variants were identified and their presence confirmed by sequencing of the Irf10 genomic region. RT-PCR showed that Irf7, Irf8 and both Irf10 transcripts were expressed in all 15 cod tissues tested, while Irf4a and Irf4b were absent in some tissues. QPCR analysis of spleen expression expanded upon this, and upon previous work. All IRF transcripts in the study were responsive to stimulation by the viral mimic poly(I:C), and all except Irf4a were responsive to exposure to formalin-killed Aeromonas salmonicida (ASAL). These IRF genes, thus, are likely important in the cod immune response to both viral and bacterial infections. Increased temperature (10 °C to 16 °C) was also observed to modulate the antibacterial responses of all IRF transcripts, and the antiviral responses of Irf4b and Irf10-v2. This research supports earlier studies which reported that elevated temperature modulates the expression of many immune genes in Atlantic cod.

摘要

干扰素调节因子(IRF)基因家族编码一组转录因子,这些转录因子不仅在调节I型干扰素(IFN)及IFN信号通路中其他基因的表达方面发挥重要作用,而且在生长、发育以及肿瘤发生调控中也具有重要作用。在本研究中,对大西洋鳕(Gadus morhua)的几个IRF家族成员(Irf4a、Irf4b、Irf7、Irf8、Irf10)进行了cDNA和推定氨基酸水平的特征分析,从而能够对硬骨鱼中的这些蛋白质进行系统发育分析,并开发用于逆转录聚合酶链反应(RT-PCR)和定量聚合酶链反应(QPCR)分析的基因特异性引物。鉴定出了两种大西洋鳕Irf10剪接变体,并通过对Irf10基因组区域进行测序证实了它们的存在。RT-PCR结果显示,Irf7、Irf8以及两种Irf10转录本在所有检测的15种鳕鱼组织中均有表达,而Irf4a和Irf4b在某些组织中不存在。对脾脏表达的QPCR分析在此基础上进一步拓展了先前的研究工作。研究中的所有IRF转录本对病毒模拟物聚肌胞苷酸(poly(I:C))的刺激均有反应,除Irf4a外,所有转录本对福尔马林灭活的杀鲑气单胞菌(ASAL)暴露也有反应。因此,这些IRF基因可能在鳕鱼对病毒和细菌感染的免疫反应中发挥重要作用。还观察到温度升高(从10℃至16℃)会调节所有IRF转录本的抗菌反应以及Irf4b和Irf10-v2的抗病毒反应。本研究支持了早期的研究报告,即温度升高会调节大西洋鳕中许多免疫基因的表达。

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